Abstract: The present invention provides an improved method for forming pH gradients or other pH functions for use in electrophoresis and isoelectric focusing. It also involves electrophoresis and IEF devices created by such methods, and improved reagents suitable for generating such devices. The method involves (1) chemically affixing or "anchoring," in a concentration gradient or other varying function, selected molecules which form acidic complexes when contacted by certain acidic compounds, and (2) contacting the anchored molecules with a fluid containing a suitable acidic compound. One example of a paired combination of anchored molecules and acid comprises (a) anchored molecules with polyhydroxyl groups and (b) boric acid, borax, and/or other borate constituents.